首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Isolation and sequence of a human apolipoprotein CII cDNA clone and its use to isolate and map to human chromosome 19 the gene for apolipoprotein CII.
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Isolation and sequence of a human apolipoprotein CII cDNA clone and its use to isolate and map to human chromosome 19 the gene for apolipoprotein CII.

机译:人载脂蛋白CII cDNA克隆的分离和序列及其在分离载人载脂蛋白CII基因并将其映射到人染色体19上的用途。

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摘要

cDNA clones encoding human apolipoprotein CII (apo CII) were identified by screening an adult human liver cDNA library with a mixed oligonucleotide probe corresponding to all possible codons for apo CII amino acid 6-10. One clone with an approximately equal to 500-base-pair (bp) insert, designated pCII -711, was selected for DNA sequence analysis. This clone contained a DNA sequence that corresponded with the previously reported amino acid sequence of apo CII with only minor differences. The DNA sequence specified a polypeptide of 79 amino acids, compared to the 78 amino acids previously reported. The pCII -711 clone contains a 36-bp DNA sequence upstream from that specifying the NH2-terminal threonine which, when read in frame, specifies the amino acid sequence Leu-Val-Leu-Leu-Val-Leu-Gly-Phe-Glu-Val-Gln-Gly and may be part of an apo CII signal peptide. The pCII -711 clone also contains a 144-bp region that corresponds to the 3' untranslated region of apo CII mRNA as well as a portion of the poly(A) tail. Clone pCII -711 was used to isolate and characterize by restriction endonuclease digestion the gene for apo CII from a human genomic library. In addition, through Southern blot analysis of DNA from human-rodent somatic cell hybrids, clone pCII -711 also was used to provisionally map the gene for apo CII to human chromosome 19.
机译:通过用混合寡核苷酸探针(对应于载脂蛋白CII氨基酸6-10的所有可能密码子)筛选成年人类肝脏cDNA文库,鉴定出编码人载脂蛋白CII(apo CII)的cDNA克隆。选择一个具有大约等于500个碱基对(bp)插入片段的克隆,称为pCII -711,用于DNA序列分析。该克隆包含的DNA序列与先前报道的载脂蛋白CII的氨基酸序列相对应,仅有很小的差异。与先前报道的78个氨基酸相比,DNA序列指定了79个氨基酸的多肽。 pCII -711克隆在指定NH2末端苏氨酸的上游含有36 bp的DNA序列,当按框读取时,该序列指定了氨基酸序列Leu-Val-Leu-Leu-Val-Leu-Gly-Phe-Glu -Val-Gln-Gly,并且可能是apo CII信号肽的一部分。 pCII -711克隆还包含一个144-bp区域,与apo CII mRNA的3'非翻译区域相对应,以及一部分poly(A)尾巴。克隆pCII -711用于通过限制性核酸内切酶消化来分离和鉴定来自人类基因组文库的载脂蛋白CII基因。另外,通过对人类啮齿动物体细胞杂种的DNA的Southern印迹分析,克隆pCII -711也被用于将载脂蛋白CII的基因临时定位于人类19号染色体。

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